Method for manufacturing substrate for making microarray

Abstract

To provide a method for manufacturing a substrate for making a microarray which will ensure the secure immobilization of a material in a site-selective manner at a low cost. The method comprises the steps of: forming a monomolecular film on the surface of a substrate using a silane compound represented by the following general formula (1), Y3Si—(CH2)m—X  (1), wherein m represents an integer from 3 to 20; X represents a hydroxyl group precursor functional group which will be converted to a hydroxyl group when exposed to acid; and Y independently represents a halogen atom or alkoxy group having 1-4 carbon atoms; and converting the hydroxyl group precursor functional group represented by X to a hydroxyl group; wherein the step of converting a hydroxyl group precursor functional group represented by X to a hydroxyl group comprises forming, on the monomolecular film, a polymer layer containing a compound represented by the following general formula (2) or (3),

The invention claimed is: 
     
       1. A method for manufacturing a substrate for making a microarray, comprising at least the steps of:
 forming a monomolecular film on the surface of a substrate using a silane compound represented by the following general formula (1),
   Y 3 Si—(CH 2 ) m —X  (1),
 
 
 wherein m represents an integer from 3 to 20; X represents a hydroxyl group precursor functional group which will be converted to a hydroxyl group when exposed to acid; and Y independently represents a halogen atom or an alkoxy group having 1-4 carbon atoms; and 
 converting the hydroxyl group precursor functional group represented by X to a hydroxyl group, wherein the step of converting the hydroxyl group precursor functional group represented by X to a hydroxyl group comprises forming, on the monomolecular film, a polymer layer containing a compound represented by the following general formula (2), 
 
       
         
           
           
               
               
           
         
         wherein R 1  may be the same or different, and represents a hydrogen atom, a fluorine atom, a chlorine atom, a nitro group, or a linear, branched, or cyclic alkyl group or alkoxy group having 1-12 carbon atoms substituted or unsubstituted; q independently represents 0 or 1; p is 1 or 2; r is an integer from 0 to 4; and r′ is an integer from 0 to 5; or a compound represented by the following general formula (3), 
       
       
         
           
           
               
               
           
         
         wherein R 2  represents a linear, branched, or cyclic alkyl group or alkoxy group having 3-10 carbon atoms substituted or unsubstituted, or a substituted or unsubstituted phenyl group or camphor; and radiating a high energy beam onto the substrate in the form of a pattern. 
       
     
     
       2. The method for manufacturing a substrate for making a microarray according to  claim 1 , wherein the hydroxyl group precursor functional group represented by X in the general formula (1) is an alkoxymethoxy group in which the alkoxy group moiety has 1-6 carbon atoms and/or an oxyranyl group. 
     
     
       3. The method for manufacturing a substrate for making a microarray according to  claim 1 , wherein the step of forming a monomolecular film using a silane compound represented by the general formula (1) comprises mixing the silane compound with at least one or more of silane compounds represented by the following general formulae (4) and (5),
   Y′ 3 Si—(CH 2 ) n —CH 3   (4)
 
   Y′ 3 Si—(CH 2 ) n —OCH 3   (5),
 
 wherein n is an integer from 0 to (m-2), m is as defined in relation to the general formula (1); and Y′ represents a halogen atom or an alkoxy group having 1-4 carbon atoms; and 
 using the resulting mixture to form the monomolecular film. 
 
     
     
       4. The method for manufacturing a substrate for making a microarray according to  claim 2 , wherein the step of forming a monomolecular film using a silane compound represented by the general formula (1) comprises mixing the silane compound with at least one or more of silane compounds represented by the following general formulae (4) and (5),
   Y′ 3 Si—(CH 2 ) n —CH 3   (4)
 
   Y′ 3 Si—(CH 2 ) n —OCH 3   (5),
 
 wherein n is an integer from 0 to (m-2), m is as defined in relation to the general formula (1); and Y′ represents a halogen atom or an alkoxy group having 1-4 carbon atoms; and 
 using the resulting mixture to form the monomolecular film. 
 
     
     
       5. The method for manufacturing a substrate for making a microarray according to  claim 1 , wherein the step of converting the hydroxyl group precursor functional group to a hydroxyl group comprises, in sequence, forming polymer layer comprising a photoacid generator on the substrate and then subjecting the substrate to heating treatment, radiating a high energy beam onto the substrate in the form of a pattern, and then subjecting the substrate to heating treatment, and then removing the polymer layer. 
     
     
       6. The method for manufacturing a substrate for making a microarray according to  claim 2 , wherein the step of converting the hydroxyl group precursor functional group to a hydroxyl group comprises, in sequence, forming polymer layer comprising a photoacid generator on the substrate and then subjecting the substrate to heating treatment, radiating a high energy beam onto the substrate in the form of a pattern, and then subjecting the substrate to heating treatment, and then removing the polymer layer. 
     
     
       7. The method for manufacturing a substrate for making a microarray according to  claim 3 , wherein the step of converting the hydroxyl group precursor functional group to a hydroxyl group comprises, in sequence, forming polymer layer comprising a photoacid generator on the substrate and then subjecting the substrate to heating treatment, radiating a high energy beam onto the substrate in the form of a pattern, and then subjecting the substrate to heating treatment, and then removing the polymer layer. 
     
     
       8. The method for manufacturing a substrate for making a microarray according to  claim 4 , wherein the step of converting the hydroxyl group precursor functional group to a hydroxyl group comprises, in sequence, forming polymer layer comprising a photoacid generator on the substrate and then subjecting the substrate to heating treatment, radiating a high energy beam onto the substrate in the form of a pattern, and then subjecting the substrate to heating treatment, and then removing the polymer layer. 
     
     
       9. The method for manufacturing a substrate for making a microarray according to  claim 1 , wherein the high energy beam has a wavelength in the range of 250 nm to 400 nm. 
     
     
       10. The method for manufacturing a substrate for making a microarray according to  claim 2 , wherein the high energy beam has a wavelength in the range of 250 nm to 400 nm. 
     
     
       11. The method for manufacturing a substrate for making a microarray according to  claim 3 , wherein the high energy beam has a wavelength in the range of 250 nm to 400 nm. 
     
     
       12. The method for manufacturing a substrate for making a microarray according to  claim 4 , wherein the high energy beam has a wavelength in the range of 250 nm to 400 nm. 
     
     
       13. The method for manufacturing a substrate for making a microarray according to  claim 1 , wherein the microarray is used for analyses of biomolecules. 
     
     
       14. The method for manufacturing a substrate for making a microarray according to  claim 2 , wherein the microarray is used for analyses of biomolecules. 
     
     
       15. The method for manufacturing a substrate for making a microarray according to  claim 3 , wherein the microarray is used for analyses of biomolecules. 
     
     
       16. The method for manufacturing a substrate for making a microarray according to  claim 4 , wherein the microarray is used for analyses of biomolecules. 
     
     
       17. The method for manufacturing a substrate for making a microarray according to  claim 1 , wherein, the method further contains a step that on the substrate in which the hydroxyl group generated through the step of converting the hydroxyl group precursor functional group to a hydroxyl group has been attached, in a dehydrated environment, a mononucleotide which has its 5′-end protected with an acid leaving group and 3′-end with phosphoramidite attached, the acid leaving group of the 5′-end is converted into a hydroxyl group using the photoacid generator contained in the polymer layer. 
     
     
       18. The method for manufacturing a substrate for making a microarray according to  claim 2 , wherein, the method further contains a step that on the substrate in which the hydroxyl group generated through the step of converting the hydroxyl group precursor functional group to a hydroxyl group has been attached, in a dehydrated environment, a mononucleotide which has its 5′-end protected with an acid leaving group and 3′-end with phosphoramidite attached, the acid leaving group of the 5′-end is converted into a hydroxyl group using the photoacid generator contained in the polymer layer. 
     
     
       19. The method for manufacturing a substrate for making a microarray according to  claim 3 , wherein, the method further contains a step that on the substrate in which the hydroxyl group generated through the step of converting the hydroxyl group precursor functional group to a hydroxyl group has been attached, in a dehydrated environment, a mononucleotide which has its 5′-end protected with an acid leaving group and 3′-end with phosphoramidite attached, the acid leaving group of the 5′-end is converted into a hydroxyl group using the photoacid generator contained in the polymer layer. 
     
     
       20. The method for manufacturing a substrate for making a microarray according to  claim 4 , wherein, the method further contains a step that on the substrate in which the hydroxyl group generated through the step of converting the hydroxyl group precursor functional group to a hydroxyl group has been attached, in a dehydrated environment, a mononucleotide which has its 5′-end protected with an acid leaving group and 3′-end with phosphoramidite attached, the acid leaving group of the 5′-end is converted into a hydroxyl group using the photoacid generator contained in the polymer layer. 
     
     
       21. The method for manufacturing a substrate for making a microarray according to  claim 17 , wherein the acid leaving group for protecting the 5′-end of the nucleotide is a dimethoxytrityl group. 
     
     
       22. The method for manufacturing a substrate for making a microarray according to  claim 18 , wherein the acid leaving group for protecting the 5′-end of the nucleotide is a dimethoxytrityl group. 
     
     
       23. The method for manufacturing a substrate for making a microarray according to  claim 19 , wherein the acid leaving group for protecting the 5′-end of the nucleotide is a dimethoxytrityl group. 
     
     
       24. The method for manufacturing a substrate for making a microarray according to  claim 20 , wherein the acid leaving group for protecting the 5′-end of the nucleotide is a dimethoxytrityl group.